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Lower NPAS3 expression during the later stages of abnormal lung development in rat congenital diaphragmatic hernia

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Abstract

Purpose

Congenital diaphragmatic hernia (CDH) is characterized by a developmental defect in the diaphragm, pulmonary hypoplasia and pulmonary hypertension. NPAS3 is a PAS domain transcription factor regulating Drosophila tracheogenesis. NPAS3 null mice develop pulmonary hypoplasia in utero and die after birth due to respiratory failure. We aimed to evaluate NPAS3 expression during normal and abnormal lung development due to CDH.

Methods

CDH was induced by administering 100 mg/ml nitrofen to time-pregnant dams on embryonic day (E) 9 of gestation. Lungs were isolated on E15, E18 and E21 and NPAS3 localization was determined by immunohistochemistry and quantified using Western blotting.

Results

We found that only E21 hypoplastic CDH lungs have reduced expression of NPAS3 in the terminal saccules. Western blotting confirmed the down-regulation of NPAS3 protein in the nitrofen-induced hypoplastic lungs.

Conclusions

We demonstrate for the first time that nitrofen-induced hypoplastic CDH lungs have reduced NPAS3 expression in the terminal saccules during the later stages of abnormal lung development. Our findings suggest that NPAS3 is associated with pulmonary hypoplasia in CDH.

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Acknowledgments

This work was supported by the Children’s Hospital Research Institute of Manitoba; RK is the recipient of a Career Enhancement Award from the Canadian Child Health Clinician Scientist Program and a New Investigator Salary Award from the Canadian Institutes of Health Research, Manitoba Lung Association and the Children’s Hospital Research Institute.

Conflict of interest

The manuscript does not contain clinical studies or patient data. The authors declare that they have no conflict of interest.

Ethical standard

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Authors and Affiliations

  1. Division of Pediatric Surgery, Departments of Surgery, Pediatrics and Child Health and Physiology and Pathophysiology (Adjunct), University of Manitoba and Children’s Hospital Research Institute of Manitoba, Biology of Breathing Theme, AE402-840 Sherbrook Street, Winnipeg, MB, R3A 1R9, Canada

    Patrícia Pereira-Terra, Ramin Kholdebarin, Meghan Higgins, Barbara M. Iwasiow & Richard Keijzer

  2. Life and Health Sciences Research Institute, 3B’s-PT Government Associate Laboratory, Braga/Guimaraes, Portugal

    Patrícia Pereira-Terra & Jorge Correia-Pinto

  3. Department of Pediatric Surgery, Hospital de Braga, Braga, Portugal

    Jorge Correia-Pinto

Authors
  1. Patrícia Pereira-Terra
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  2. Ramin Kholdebarin
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  3. Meghan Higgins
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  4. Barbara M. Iwasiow
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  5. Jorge Correia-Pinto
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  6. Richard Keijzer
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Corresponding author

Correspondence to Richard Keijzer.

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Pereira-Terra, P., Kholdebarin, R., Higgins, M. et al. Lower NPAS3 expression during the later stages of abnormal lung development in rat congenital diaphragmatic hernia. Pediatr Surg Int 31, 659–663 (2015). https://doi.org/10.1007/s00383-015-3703-2

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  • DOI: https://doi.org/10.1007/s00383-015-3703-2

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